Why is actin used as a loading control?
Beta-Actin (42 kDa) is commonly chosen as a loading control due to its general expression across all eukaryotic cell types. The expression levels of this protein do not vary drastically due to cellular treatment, which is another reason the protein makes a suitable control.
What is a loading control antibody?
Loading control antibodies are important controls as they indicate the equal loading of samples across all wells. Loading controls also indicate the proper transfer of proteins to the membrane during the western blotting process. Loading controls are typically proteins with high and ubiquitous expression.
Is alpha tubulin a loading control?
Alpha-, beta-, and gamma-tubulins have all been used as loading controls. Figure 4. Tubulins as the structural components of microtubules and the targets of drugs.
What is the point of a loading control?
Loading controls are antibodies that are used to detect proteins within samples. When western blots are used to determine the levels of protein expression in a sample, loading controls ensure that the results aren’t due to loading or protein transfer errors.
What is beta actin used for?
They are also used as loading controls in protein assays. Beta-actin is known as a “housekeeping” protein, i.e. it is expressed constantly, and at high levels, in all the cell types used in protein research. It is therefore a useful loading control in, for example, Western blot analysis.
What is a loading control in northern blot?
The election of the correct loading control in Northern blot normalization is something essential to obtain valid results. Housekeeping genes are widely used as loading control and the assumption is made that they counteract load differences between samples.
Does a loading control show normalization?
A loading control protein confirms equal sample loading and gel-to-membrane transfer. Normalized expression is then compared across samples to confirm that changes in protein of interest expression represent real differences and are not the result of differences in total protein abundance from sample-to-sample.
Why is tubulin used as a control?
Beta-Tubulin, is usually used as loading control for Western Blot to normalize the levels of protein detected by confirming that protein loading is the same across the gel.
What are alpha and beta tubulin?
Alpha tubulin refers to one of the two protein components of a microtubule attached to GTP while beta tubulin is the second protein component of a microtubule. Thus, this is the fundamental difference between alpha and beta tubulin.
Is actin a good loading control?
Beta-Actin Protein Beta-actin, also known as a “housekeeping” protein, is usually used as a loading control , for among others, the integrity of cells, protein degradation, in PCR and Western blotting. Actins are highly conserved proteins that are involved in cell motility, structure, and integrity.
Can a beta actin be used as a loading control?
Both beta and alpha actins have been used as loading controls in Western blot experiments. Please see the detailed discussion about beta-actin loading control and commonly used clones, and frequently asked questions about beta-actin controls.
How is human beta actin homologous to other actins?
Human beta actin shares close to 90% identity with fungal homologs. Human beta actin is at least 93% identical with other members of the human actin family. This homology among actins and across species is important for antibody selection and the interpretation of Western blot bands.
What is the function of actin in cells?
Actin: an essential player in cell adhesion and migration. Found in all eukaryotic cells, actin is a protein essential to maintain cell structure. In humans, three main actin groups have been identified – alpha, beta, and gamma. The alpha actins are expressed primarily in skeletal, cardiac, and smooth muscle, while the remaining beta and gamma.
What are the function of alpha actin filaments?
The alpha actins are expressed primarily in skeletal, cardiac, and smooth muscle, while the remaining beta and gamma. Actin filaments provide internal mechanical support to maintain cell shape, as well as the force to drive cell movements and intracellular transport.